Optimizing gamma-ray spectrometers for UAV-borne surveys with geophysical applications.

Heavy duty unmanned aerial vehicles (UAVs) have made it possible to fly with large gamma-ray spectrometers that weigh several kilograms. Moreover, they can be purchased at an affordable price. These large UAV-borne gamma-ray detection systems are used to map the naturally occurring radionuclides 40K, 238U, 232Th. Such platforms have the advantage that they can be deployed over terrain that is difficult to access, while still maintaining a high spatial resolution. In contrast to UAV-borne radioactive pollution studies, the naturally occurring radionuclides have a much lower activity and therefore require longer integration time, slower flying speed or a larger detector, in order to effectively determine the spatial radionuclide distribution. Therefore, the question arises: what is the minimum practical detector size required to successfully map 40K, 238U and 232Th concentrations from UAV platforms. In this study an agricultural field has been mapped with three different scintillator-based gamma-ray spectrometers: a 2000 ml, 1000 ml, and 350 ml detector. They were mounted together on the same UAV. At a flying height of 20 m and a speed of 5.6 m s-1 the field was mapped. The various aerial measurements were compared to each other and to the ground-based measurements. The field had a low spatial variation in the 40K concentration (relative standard deviation (RSD) = 9%) and a larger variation for 238U and 232Th concentrations (RSD = 24% and 31% respectively). Radionuclide concentrations have been extracted from the survey data by Full Spectrum Analysis (FSA). Uncertainties and variances of the radionuclides have been determined by using two methods. Firstly, they are calculated directly from the FSA output and secondly they are extracted from a variogram. The latter incorporates spatial variation and was shown to provide a lower uncertainty. When using small detectors, the former approach could lead to the conclusion that the uncertainty is larger than the variance, while the variogram approach does capture the spatial variation. All three detectors were able to characterize the spatial distribution of the 232Th concentration. It is shown that the 232Th concentration is a good predictor of the sand and clay fraction of the topsoil in the field. By comparing the UAV-borne measurements to the ground-based measurements it is found that UAV-borne measurements at 20 m height are less sensitive to extreme values than ground-based measurements and they have the tendency to shift to the mean concentration of the area. The results of this study can be used to optimize the detector volume, survey height, and survey speed to maintain an acceptable accuracy for gamma-ray studies with small UAV-borne detectors.

Outcome after Hybrid Outflow Interventions for Chronic Limb-threatening Ischemia.

BACKGROUND AND AIMS: Because chronic limb-threatening ischemia (CLTI) is often associated with multilevel arterial disease, it usually requires revascularization at different sites of the limb vasculature. We aim to assess the outcome of the hybrid interventions including open surgical revascularization together with outflow segment percutaneous transluminal angioplasty (PTA) in patients with chronic limb-threatening ischemia. MATERIAL AND METHODS: This study included all hybrid outflow-PTA interventions (n = 80) on patients suffering from CLTI performed in Helsinki University Hospital between 2003 and 2015. Follow-up ended on 31 December 2019. Patient data were prospectively collected into our vascular registry and scrutinized retrospectively. Thirty-one patients (39%) suffered from rest pain (Rutherford category IV) and 49 patients (61%) had ischemic ulcers (Rutherford category V-VI). The most common open surgical procedure was femoral endarterectomy (n = 63, 79%) and the most common endovascular procedure was superficial femoral artery percutaneous transluminal angioplasty (n = 65, 81%). Mean follow-up time was 56 months (range: 4 days-183 months). RESULTS: Limb salvage was at 30 days-92%, at 1 year-91%, and at 5 and 10 years-86%. Survival and amputation-free survival were at 30 days-93% and 86%, at 1 year-80% and 76%, at 5 years-51% and 48%, and at 10 years-21% and 21%. Wound healing at 3, 6, and 12 months was 48%, 71%, and 87%. Freedom from target lesion revascularization was at 30 days-97%, at 1 year-88%, at 5 years-72%, and at 10 years-66%. CONCLUSION: Hybrid outflow revascularization is an important tool in the vascular surgeon's armamentarium for treatment of patients with multilevel arterial disease causing chronic limb-threatening ischemia.

Long-term Results of Inframalleolar Bypass for Critical Limb Ischaemia.

INTRODUCTION: As the population ages and the incidence of diabetes increases, the expected number of patients with critical limb ischaemia (CLI) requiring distal revascularization will remain high or even increase. The aim of this study was to investigate the long-term results of inframalleolar bypass. MATERIAL AND METHODS: A total of 352 inframalleolar bypasses for CLI performed between 2002 and 2013 were included. Risk factors were evaluated and patency (both clinical and imaging based), leg salvage, survival, and amputation free survival (AFS) assessed. RESULTS: The median follow up was 30 months (mean 42 months, range 1-186 months). The median age of the study population was 73 years, and 67% of the patients were male. The incidence of diabetes was 69%. In the majority of cases (82%), the indication for bypass was an ulcer or gangrene, and the remaining 18% of the patients had rest pain. Primary, assisted primary, and secondary clinical patency was 71.2%, 76.5%, 81.0%, and 59.7%, 69.3%, and 70.7%, and 49.0%, 58.6%, and 68.4% at 1, 5, and 10 years, respectively. The last imaging based secondary patency at 1, 5, and 10 years was 79.3%, 68.1%, and 62.8%, respectively. The popliteal artery as the inflow artery (n = 194) was associated with superior primary (p = .013), assisted primary (p = .028), and secondary patency (p = .014) when compared with bypasses originating from the femoral artery (n = 158). The leg salvage rate at 1, 5, and 10 years was 78.6%, 72.0%, and 67.2%, respectively. Leg salvage was equal in patients with and without diabetes (p = .460). The respective survival and AFS rates at 1, 5, and 10 years were 70.3%, 37.4%, and 15.9%, and 58.4%, 29.8%, and 12.8%. CONCLUSION: Bypass to the foot arteries yielded excellent long-term patency, and good limb salvage can be achieved in both non-diabetic and diabetic patients.

Differential impact of bypass surgery and angioplasty on angiosome-targeted infrapopliteal revascularization.

OBJECTIVE: The aim of this study was to evaluate the impact of angiosome targeted revascularization according to the revascularization method. DESIGN: Retrospective observational study. MATERIALS AND METHODS: This study cohort comprised 744 consecutive patients who underwent infrapopliteal endovascular or surgical revascularization between January 2010 and July 2013. Differences in outcomes after bypass surgery and PTA were adjusted by estimating a propensity score, which was employed for one to one matching as well as adjusted analysis. RESULTS: Cox proportional hazards analysis showed that angiosome-targeted revascularization (HR 1.29, 95% CI 1.02-1.65), bypass surgery (HR 1.79, 95% CI 1.41-2.27), C-reactive protein ≤10 mg/dL (HR 1.42, 95% CI 1.11-1.81), and the number of affected angiosomes (HR 0.85, 95% CI 0.74-0.98) were independent predictors of improved wound healing. When adjusted for the number of affected angiosomes and C-reactive protein ≤10 mg/dL, angiosome-targeted bypass surgery was associated with a significantly higher rate of wound healing than non-angiosome-targeted angioplasty (HR 2.27, 95% CI 1.61-3.20). This was confirmed in propensity score adjusted analysis (HR 1.72, 95% CI 1.35-2.16). Among patients who underwent angiosome-targeted revascularization, the propensity score adjusted analysis showed that bypass surgery was associated with a significantly better rate of wound healing (HR 154, 95% CI 1.09-2.16) but similar limb salvage rates when compared with angioplasty (HR 0.79, 95% CI 0.44-1.43). CONCLUSION: Rates of wound healing and limb salvage in patients with critical limb ischemia (CLI) were significantly better after angiosome-targeted revascularization, bypass surgery achieving significantly better wound healing than angioplasty.

Cdc20 and securin overexpression predict short-term breast cancer survival.

BACKGROUND: Cdc20 is an essential component of cell division and responsible for anaphase initiation regulated by securin degradation. Cdc20 function is strongly regulated by the spindle assembly checkpoint to ensure the timely separation of sister chromatids and integrity of the genome. We present the first results on Cdc20 in a large clinical breast cancer material. METHODS: The study was based on 445 breast cancer patients with up to 20 years of follow-up (mean 10.0 years). DNA content was determined by image cytometry on cell imprints, and Cdc20 and securin immunohistochemistry on tissue microarrays of breast cancer tissue. RESULTS: In our results, high Cdc20 and securin expression was associated with aneuploid DNA content. In prognostic analyses, high Cdc20 immunoexpression alone and in combination with high securin immunoexpression indicated aggressive course of disease and up to 6.8-fold (P<0.001) risk of breast cancer death. Particularly, high Cdc20 and securin immunoexpression identified a patient subgroup with extremely short, on average 2.4 years, breast cancer survival and triple-negative breast cancer (TNBC) subtype. CONCLUSIONS: We report for the first time the association of high Cdc20 and securin immunoexpression with extremely poor outcome of breast cancer patients. Our experience indicates that Cdc20 and securin are promising candidates for clinical applications in breast cancer prognostication, especially in the challenging prognostic decisions of TNBC.

Angiosome theory: fact or fiction?

The angiosome concept delineates the human body into three-dimensional blocks of tissue fed by specific arterial and venous sources named "angiosomes." Adjacent angiosomes are connected by a vast compensatory collateral web, or "choke vessels." This concept may provide new information applicable to improving targeted revascularization of ischemic tissue lesions. A few dedicated studies available seem to favor this strategy, as encouraging ulcer healing and limb preservation are reported in connection with both bypass and endovascular techniques based on these principles. The theory on the angiosome model of revascularization (AMV) may help the clinician to better refine vessel selection, vascular access, and specific strategies in the revascularization of critically ischemic legs with tissue lesions. Specific applications of angiosome-guided revascularization were recently suggested for patients with diabetes or renal insufficiency, with ischemic tissue lesions of the lower limb, and extended large- and medium-size collateral network decay. For these cases, the concept may allow deliberate arterial reconstruction following individual wound topographies in specific ischemic areas, although deprived from "rescue-vessel" supply. The AMV theory may contribute to a shift in common reperfusion options. However, the data available is suggestive and does not provide strong evidence as factors such as case mix and the severity of ischemia are unsatisfactorily controlled. At present, the evidence is scarce as to the effect of the severity of the arterial disease. In all comparisons, the groups treated are likely to be dissimilar and mismatched. The angiosome concept is postulated to be valid especially in diabetics, whose ischemic lesions tend to heal worse than those of non-diabetics.

End-stage renal disease and critical limb ischemia: a deadly combination?

OBJECTIVES: This study was planned to evaluate the prognostic impact of end-stage renal disease (ESRD) in patients with critical leg ischemia (CLI) undergoing infrainguinal revascularization. MATERIALS AND METHODS: 1425 patients who underwent infrainguinal revascularization for CLI were the subjects of the present analysis. Ninety-five patients had ESRD (eGFR < 15 ml/min/m²), and of them 66 (70%) underwent percutaneous transluminal angioplasty and 29 (30%) underwent bypass surgery. RESULTS: ESRD patients had significantly lower overall survival (at 3-year, 27.1% vs. 59.7%, p < 0.0001), leg salvage (at 3-year, 57.7% vs. 83.0%, p < 0.0001), and amputation free survival (at 3-year, 16.2% vs. 52.9%, p < 0.0001) than patients with no or less severe renal failure. The difference in survival was even greater between 86 one-to-one propensity matched pairs (at 3-year, 23.1% vs. 67.3%, p < 0.0001). ESRD was an independent predictor of all-cause mortality (RR 2.46, 95%CI 1.85-3.26). Logistic regression showed that age ≥ 75 years was the only independent predictor of 1-year all-cause mortality (OR 4.92, 95%CI 1.32-18.36). Classification and regression tree analysis showed that age ≥ 75 years and, among younger patients, bypass surgery for leg ulcer and gangrene were associated with significantly higher 1-year mortality CONCLUSIONS: Lower limb revascularization in patients with CLI and end-stage renal failure is associated with favourable leg salvage. However, these patients have a very poor survival and this may jeopardize any attempt of revascularization. Further studies are needed to identify ESRD patients with acceptable life expectancy and who may benefit from lower limb revascularization.

The role of chronic kidney disease as a predictor of outcome after revascularisation of the ulcerated diabetic foot.

AIMS/HYPOTHESIS: The aim of the study was to stratify the risk of diabetic patients with leg ulcer or gangrene undergoing infrainguinal revascularisation for critical limb ischaemia. METHODS: The study cohort included 732 revascularisation procedures performed in 597 diabetic patients with ulcer or gangrene. Logistic regression and CART analysis were used for identification of predictors of 1-year outcome. RESULTS: Logistic regression showed that chronic kidney disease (CKD) class (OR 1.38, 95% CI 1.16, 1.65) was an independent predictor of 1-year leg salvage (area under the receiver operating characteristic [ROC] curve 0.60, 95% CI 0.54, 0.65). The terminal nodes of the CART for 1-year leg salvage were CKD classes 4-5, the level (infrapopliteal vs femoropopliteal revascularisation), type of revascularisation (bypass surgery vs percutaneous transluminal angioplasty) and gangrene (area under the ROC curve 0.62, 95% CI 0.57, 0.68). Logistic regression showed that pulmonary disease (OR 1.76, 95% CI 1.11, 2.78), CKD class (OR 1.43, 95% CI 1.24, 1.65), foot gangrene (OR 1.76, 95% CI 1.21, 2.60) and patient age (OR 1.02, 95% CI 1.01, 1.04) were independent predictors of 1-year amputation-free survival (area under the ROC curve 0.65, 95% CI 0.60, 0.69). The terminal nodes of the CART for 1-year amputation-free survival were CKD classes 3-5, patient's age of ≥ 75 years and foot gangrene (area under the ROC curve 0.64, 95% CI 0.60, 0.68). CONCLUSIONS/INTERPRETATION: CKD is a formidable risk factor for poor intermediate outcome after infrainguinal revascularisation in diabetic patients with foot ulcer or gangrene. CART analysis indicates that foot gangrene is also a significant risk factor for adverse outcome.

Infrainguinal percutaneous transluminal angioplasty or bypass surgery in patients aged 80 years and older with critical leg ischaemia.

BACKGROUND: Infrainguinal revascularization for critical leg ischaemia (CLI) in patients aged 80 years and over is associated with increased operative risk. The aim was to compare the results of percutaneous transluminal angioplasty (PTA) and bypass surgery in these patients. METHODS: Some 584 consecutive patients aged at least 80 years treated with either PTA (277) or bypass surgery (307) for CLI between 2000 and 2007 were included in this study. RESULTS: After 2 years PTA achieved better results than bypass surgery (leg salvage: 85.4 versus 78.7 per cent, P = 0.039; survival: 57.7 versus 52.3 per cent, P = 0.014; amputation-free survival (AFS): 53.0 versus 44.9 per cent, P = 0.005). Cox regression analysis showed that increased age (relative risk (RR) 1.05, 95 per cent confidence interval 1.02 to 1.08), decreased estimated glomerular filtration rate (RR 0.99, 0.99 to 1.00), diabetes (RR 1.30, 1.04 to 1.62), coronary artery disease (RR 1.36, 1.05 to 1.75) and bypass surgery (RR 1.55, 1.24 to 1.93) were associated with decreased AFS. In 95 propensity score-matched pairs, leg salvage at 2 years (88 versus 75 per cent; P = 0.010) and AFS (53 versus 45 per cent; P = 0.033) were significantly better after PTA. Classification and regression tree analysis suggested that PTA was associated with better 1-year AFS, especially in patients with coronary artery disease (63.8 versus 48.9 per cent; P = 0.008). CONCLUSION: When feasible, a strategy of PTA first appears to achieve better results than infrainguinal bypass surgery in patients aged 80 years and older.

Femoropopliteal balloon angioplasty vs. bypass surgery for CLI: a propensity score analysis.

OBJECTIVES: To compare the outcomes of femoropopliteal percutaneous transluminal angioplasty (PTA) and bypass surgery for critical limb ischaemia (CLI). DESIGN: The study is retrospective in nature. MATERIALS AND METHODS: This study included 858 consecutive patients, who underwent femoropopliteal revascularisation for CLI at Helsinki University Central Hospital during 2000-2007. As many as 517 patients (60%) underwent PTA and 341 (40%) bypass surgery. Propensity score analysis was used for risk adjustment in multivariable analysis and for one-to-one matching. RESULTS: In the overall series, PTA had poorer long-term results than bypass (5-year leg salvage, 78.2% vs. 91.8%, p < 0.0001; survival 49.2% vs. 57.1%, p = 0.048; amputation-free survival, 42.0% vs. 53.7%, p = 0.003; freedom from surgical re-intervention 86.2% vs. 94.3%, p < 0.0001). When treatment method was adjusted for propensity score as well as in the propensity score-matched pairs, leg salvage and freedom from surgical re-intervention were worse after PTA than after bypass (among the 241 propensity score-matched pairs, 74.3% vs. 88.2%, p = 0.031, and 86.1% vs. 89.8%, p = 0.025, respectively). Differences in survival, amputation-free survival and freedom from any re-intervention were not observed. CONCLUSIONS: In CLI patients, femoropopliteal PTA seems to be associated with poorer long-term leg salvage and freedom from surgical re-intervention than bypass surgery. However, the treatment method did not affect long-term amputation-free survival.

High leg salvage rate after infrainguinal bypass surgery for ischemic tissue loss (Fontaine IV) is compromised by the short life expectancy.

BACKGROUND AND AIMS: Most studies analysing the prognosis of infrainguinal bypass surgery (IBS) in patients with critical leg ischemia (CLI) have combined the outcome of patients with rest pain and tissue loss. The aim of the present study was to evaluate amputation-free survival (AFS) after IBS in patients with the most advanced form of peripheral arterial disease, CLI with tissue loss (Fontaine IV), and to analyse the risk factors for an adverse outcome. PATIENTS AND METHODS: 636 patients with CLI and tissue loss who underwent unilateral IBS between January 2000 and December 2006 at our institution were included in this retrospective study. RESULTS: At one year, the leg salvage, survival and amputation-free survival rates were 83%, 71% and 55%, respectively, and at five years 76%, 38% and 30%, respectively. In univariate analysis, diabetes was associated with decreased AFS. In multivariate analysis, age, coronary artery disease, chronic pulmonary disease, gangrene and renal insufficiency were independent risk factors for decreased AFS. CONCLUSION: Infrainguinal bypass grafting results in a high rate of leg salvage. Amputation-free survival was low during the follow-up due to the high mortality of patients with CLI and tissue loss. Several co-morbidities of the CLI patients were associated with decreased amputation-free survival.

Healing of ischaemic tissue lesions after infrainguinal bypass surgery for critical leg ischaemia.

OBJECTIVE: To evaluate healing time of ischaemic tissue lesions, limb salvage and survival in patients suffering from critical limb ischaemia (CLI) with tissue loss. DESIGN: Prospective single centre cohort study. MATERIAL AND METHODS: Consecutive patients with CLI and tissue loss (Fontaine IV) (148 patients, 150 limbs) were followed prospectively for 1 year after infrainguinal bypass. Healing time of tissue lesions, graft patency, limb salvage, survival rates and the overall need for any type of surgical and endovascular procedure were analysed. Patient comorbidities were assessed by uni- and multivariate analysis to determine risk factors for adverse outcome. RESULTS: Complete tissue healing, including healing of ischaemic tissue lesions and surgical wounds, at 6 and 12 months after the infrainguinal bypass were respectively 40% and 75%. The median time to complete tissue healing was 190 days. Diabetes was the only significant risk factor which delayed tissue healing. Overall patency, limb salvage, survival and amputation-free survival rates were respectively at 12 months 80%, 81%, 73% and 63%. The clinically important endpoint amputation-free survival with completely healed wounds was attained in 50% of patients at 1 year. CONCLUSION: Complete healing of ischemic tissue lesions is slow even after a successful infrainguinal bypass.

Estimated glomerular filtration rate (eGFR) as a predictor of outcome after infrainguinal bypass in patients with critical limb ischemia.

OBJECTIVES: Renal insufficiency is a risk factor for poor outcome after infrainguinal bypass in patients with critical limb ischemia (CLI). Estimated glomerular filtration rate (eGFR) takes age, gender and body size into account and therefore represents actual renal function more accurately than serum creatinine level alone. The aim of this study was to determine the impact of different stages of renal insufficiency on outcome and to assess the prognostic significance of eGFR in patients with CLI. MATERIAL AND METHODS: 603 patients with CLI who underwent infrainguinal bypass between January 2002 and December 2005 at our institution were included in this retrospective study. We estimated GFR using the Modification of Diet in Renal Disease (MDRD) Study equation. Survival, leg salvage and amputation-free survival were calculated using Kaplan-Meier method. Cox regression analysis was performed to calculate hazard ratios for different outcome variables. RESULTS: Adjusted hazard ratio (HR) of mortality, limb loss and limb loss and/or death for eGFR < 30 ml/min/1.73 m(2) versus serum creatinine > 200 micromol/l was 4.0 (95% CI 2.22-7.39) vs 3.5 (95% CI 1.82-6.84), 6.5 (95% CI 2.71-15.59) vs 6.2 (95% CI 2.47-15.56) and 4.0 (95% CI 2.40-6.63) vs 3.6 (95% CI 2.03-6.25), respectively. CONCLUSION: Estimated GFR is better predictor of survival, leg salvage and amputation-free survival than serum creatinine alone. eGFR < 30 ml/min/1.73 m(2) is independent risk factor for all three outcome endpoints.

Multiple sclerosis-associated retrovirus and optic neuritis.

One prognostic factor for early multiple sclerosis (MS) patients to develop a definite MS may be the presence of the MS-associated retrovirus (MSRV) in the cerebrospinal fluid (CSF). We designed a specific study on a cohort of optic neuritis (ON) patients to evaluate the MSRV-dependent conversion to MS relative to the prediction conferred by magnetic resonance imaging (MRI) and CSF abnormalities. At follow-up, 33.3% MSRV+ and 0% MSRV- ON patients developed MS (P = 0.03). The prediction value is lower than that given by CSF and MRI abnormalities (42.3%). This intriguing finding is discussed in the light of the abundant discrepancies observed in the MSRV literature. Multiple Sclerosis 2006; 12: 357-359. www.multiplesclerosisjournal.com

Up regulation of cathepsin K expression in articular chondrocytes in a transgenic mouse model for osteoarthritis.

OBJECTIVES: To study the expression of cysteine proteinases, particularly cathepsin K, and their extracellular inhibitor cystatin C in articular cartilage of transgenic Del1 mice which harbour a short deletion mutation in a type II collagen transgene and are predisposed to early onset osteoarthritis. METHODS: Northern analysis was used to measure mRNA levels of cathepsins B, H, K, L, and S, and cystatin C in total RNA extracted from knee joints of Del1 mice, using their non-transgenic litter mates as controls. Immunohistochemistry and morphometry was used to study the distribution of cathepsin K and cystatin C in the knee joints. RESULTS: Up regulation of cathepsin K mRNA expression was seen in the knee joints of transgenic Del1 mice at the onset of cartilage degeneration. Cathepsin K was found near sites of matrix destruction in articular chondrocytes, particularly in clusters of proliferating cells, and in calcified cartilaginous matrix. In intact articular cartilage of control animals, cathepsin K was only seen in a small number of chondrocytes. Upon aging, control animals also developed osteoarthritis, which was accompanied by increased cathepsin K expression. Cystatin C was mostly localised in and around chondrocytes located in calcified cartilage, with no obvious association with the onset of cartilage degeneration. CONCLUSION: The temporospatial distribution of cathepsin K in osteoarthritic cartilage suggests a role for this enzyme in the pathogenesis of osteoarthritis. Because cathepsin K can digest cartilage matrix components it may contribute to the development of osteoarthritic lesions. These data may provide new clues for the development of treatments aimed at preventing cartilage degeneration.

Differential recruitment of the coactivator proteins CREB-binding protein and steroid receptor coactivator-1 to peroxisome proliferator-activated receptor gamma/9-cis-retinoic acid receptor heterodimers by ligands present in oxidized low-density lipoprotein.

Peroxisome proliferator-activated receptor gamma (PPARgamma) colocalizes with oxidized low-density lipoprotein (LDL) in foam cells in atherosclerotic lesions. We have explored a potential role of oxidized fatty acids in LDL as PPARgamma activators. LDL from patients suffering from intermittent claudication due to atherosclerosis was analyzed using HPLC and gas chromatography/mass spectrophotometry and found to contain 9-hydroxy and 13-hydroxyoctadecadienoic acid (9- and 13-HODE), as well as 5-hydroxy-, 12-hydroxy- and 15-hydroxyeicosatetraenoic acid (5-, 12- and 15-HETE respectively). PPARgamma was potently activated by 13(S)-HODE and 15(S)-HETE, as judged by transient transfection assays in macrophages or CV-1 cells. 5(S)- and 12(S)-HETE as well as 15-deoxy-Delta(12,14)-prostaglandin J(2) also activated PPARgamma but were less potent. Interestingly, the effect of the lipoxygenase products 13(S)-HODE and 15(S)-HETE as well as of the drug rosiglitazone were preferentially enhanced by the coactivator CREB-binding protein, whereas the effect of the cyclooxygenase product 15-deoxy-Delta(12,14)-prostaglandin J(2) was preferentially enhanced by steroid receptor coactivator-1. We interpret these results, which may have relevance to the pathogenesis of atherosclerosis, to indicate that the lipoxygenase products on the one hand and the cyclooxygenase product on the other exert specific effects on the transcription of target genes through differential coactivator recruitment by PPARgamma/9-cis retinoic acid receptor heterodimer complexes.

Cerebrospinal fluid affects phenotype and functions of myeloid dendritic cells.

Myeloid (CD11c+) dendritic cells (DC) are present in cerebrospinal fluid (CSF), as well as in the meninges and choroid plexus. Functional studies of these DC are hindered or impossible. To obviate this problem, we investigated the effects of CSF supernatants from patients with non-inflammatory neurological diseases (NIND), multiple sclerosis (MS), bacterial meningitis (BM) and Lyme meningoencephalitis (LM) on immature monocyte-derived DC (moDC) from healthy donors. CSF supernatants caused maturation of moDC (MS > LM > NIND > BM), as reflected by a decrease in CD1a, and an increase in HLA-DR, CD80 and CD86 expression. The maturation effect of MS CSF and LM CSF could be blocked by anti-TNF-alpha MoAb or recombinant human IL-10. moDC cultured with BM CSF either remained immature or turned into CD14+ macrophage-like cells and were relatively inefficient at inducing T cell responses in vitro. In contrast, moDC cultured with LM CSF induced strong Th1 responses. Both BM CSF and LM CSF contained IFN-gamma, a cytokine that augments IL-12 production by moDC and hence should confer an ability to induce a Th1 response. However, BM CSF also contained high levels of IL-10, which could antagonize the effects of IFN-gamma on moDC. moDC cultured with MS CSF induced a higher production of IFN-gamma from T cells compared to moDC cultured with NIND CSF or BM CSF. In summary, soluble factors present in the CSF may influence the phenotype and functions of meningeal, choroid plexus and CSF DC which, in turn, may have an impact on the character of intrathecal T cell responses.

Glatiramer acetate and IFN-beta act on dendritic cells in multiple sclerosis.

Glatiramer acetate (GA; Copolymer 1; Copaxone) and interferon-beta (IFN-beta) modulate the course of multiple sclerosis (MS), but probably by different mechanisms. GA, a mixture of synthetic peptides, is believed to act as an altered peptide ligand with inhibitory effects on autoreactive T cells and promoting Th2 cells. It is unknown whether GA affects dendritic cells (DCs), which, besides strong antigen presenting capacity, orchestrate Th1 and Th2 responses. IFN-beta inhibits IL-12 production by DCs over unknown mechanisms. This study was designed to investigate in vitro effects of GA and IFN-beta on the development and function of DCs from MS patients and healthy controls, and to explore their possible synergistic effects on DCs. DCs were generated from adherent blood mononuclear cells (MNCs). GA or IFN-beta or both, when added at initiation of DC cultures, rapidly promoted the development of adherent MNCs into floating, activated DCs as reflected by up-regulation of HLA-DR and CD86 expression. IFN-beta, but not GA, induced IL-3R expression on DCs. Compared to DCs from healthy controls, MS patients' DCs expressed higher levels of the myeloid DC marker CD1a and produced lower amounts of IL-10. GA reduced IL-12 production by DCs. IFN-beta also reduced IL-12, but increased IL-10 production by DCs from both MS patients and healthy controls. GA and IFN-beta synergistically suppressed CD1a and enhanced CD86 expression on MS DCs. These findings document novel mechanisms of action of GA and IFN-beta at the DC level in MS.

Multiple sclerosis: deficient in vitro responses of blood mononuclear cells to IFN-beta.

IFN-beta may modify the clinical course of multiple sclerosis (MS) but is not curative, and there are also patients whose disease does not respond to IFN-beta as currently administered. Tests are warranted with a capacity to early discriminate responders from non-responders, thereby altering treatment option for the individual patient. In vitro effects of IFN-beta on expression of activation-associated cell surface markers and cytokine production need to be explored in this context. Here we report on the influence in vitro of IFN-beta on blood mononuclear cells (MNC) prepared from MS patients and healthy controls. MNC were subjected to short-term culture in the presence of IFN-beta at concentrations of 100 U/ml and 1000 U/ml. Expression of cell surface molecules CD40, CD69, CD80, CD86, CD95 and HLA-DR was measured by flow cytometry. IL-10 and IL-12 p40 production in culture supernatants was measured by ELISA. MNC exposed to IFN-beta in vitro enhanced expression of the co-stimulatory CD80, CD86, the early activation antigen CD69 and the cell death receptor CD95. Expression of CD40 and HLA-DR was not influenced. IFN-beta increased IL-10 but suppressed IL-12 p40 production. In vitro effects of IFN-beta on MNC were similar in MS patients and in healthy subjects, except that IFN-beta-induced augmentation of CD86 and CD69 expression was less pronounced in MS, in particular in untreated MS patients. Individual MS patients clearly responded differently to IFN-beta in vitro in comparison with the majority of patients in this cross-sectional study. In conclusion, anti-inflammatory effects of IFN-beta on blood MNC include augmentation of IL-10 production and suppression of IL-12 p40 production, which are accompanied by enhancement of CD69, CD80, CD86 and CD95 expression. The less pronounced IFN-beta-induced effects on CD86 and CD69 expression in MS vs controls might reflect a defect in immunoregulation in MS. Larger groups should be evaluated, and follow-up studies performed in MS patients before/during IFN-beta treatment in relation to clinical outcome measures to evaluate the usefulness of these markers for possible differentiation between responders and non-responders to IFN-beta treatment.

Expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in human chondrosarcomas.

The aim of the present study was to characterise the ability of malignant chondrosarcomas to invade normal bone by analysing their production of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). For this purpose 12 chondrosarcomas were investigated for the expression of mRNAs for several MMPs and all 4 TIMPs by Northern hybridisation, and for immunohistochemical localisation of the proteins. A characteristic finding of these analyses was increased expression of MMP-13, MMP-14 and TIMP-2 mRNAs in chondrosarcomas when compared with nonmalignant control samples. Individual chondrosarcomas also exhibited elevated levels of MMP-1, MMP-7 and MMP-9 mRNAs. The results of Northern hybridisations were supported by immunohistochemical stainings of the corresponding tumour areas for MMP-2, MMP-14 and TIMP-2, further suggesting that these may have prognostic value for determining whether individual chondrosarcomas are locally aggressive or have a probability of recurrence. Another finding of the present study was a marked heterogeneity in histologic appearance and gene expression of the chondrosarcomas, emphasising the importance of analysing several areas of these tumours to get representative results. These findings suggest that analysis of MMPs could be a useful diagnostic indicator in patients with cartilaginous tumours and could help in differentiating between a low-grade malignant chondrosarcoma and a benign growing enchondroma.